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Cells were resuspended in 200 l staining buffer and analyzed within the hour using a CytoFLEX S flow cytometer (Beckman Coulter)

Basal insulin was measured in plasma from female [Young ( n = 16), Old Con ( n = 26), and Old mAb ( n = 25)] and male mice [Young ( n = 17), Old Con ( n = 32), and Old mAb ( n = 28)] using a bead-based assay for mouse insulin (EMD Millipore, Inc) and detection was performed on a Bio-Plex MAGPIX Multiplex Reader (Biorad Inc., Hercules, CA)

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